LOCUS       V043999                 8306 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043999
VERSION     V043999
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..3898
                     /label="Atp2a2(NM_001110139)"
                     /note="Atp2a2(NM_001110139)"
                     /gene="Atp2a2"
     CDS             3932..4021
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    4068..4292
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4338..4766
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4780..5109
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5176..5775
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5949..6070
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6119..6135)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6143..6159
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6167..6197)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6212..6233
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6521..7106)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7277..8137)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8138..8242)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gagaacgctc acacaaagac cgtggaggag gtgctgggcc acttcggcgt
      961 caacgagagc acggggctga gcctggagca ggtcaagaag ctcaaggaga gatggggctc
     1021 caacgaattg ccggctgaag aaggaaagac cttgctggaa cttgtgatcg agcagtttga
     1081 agacttacta gttagaattt tattgctggc agcatgtata tctttcgttt tggcttggtt
     1141 cgaagaaggt gaagaaacga ttacagcctt tgtagaacct tttgtaattc tgcttatatt
     1201 ggtagccaat gcaattgtgg gtgtatggca ggagagaaac gctgaaaatg caatagaagc
     1261 tcttaaggag tatgaacctg aaatgggcaa ggtgtatcga caggacagaa agagtgtgca
     1321 gcggattaaa gcgaaagata tagttcctgg ggatatagtg gaaattgctg ttggtgacaa
     1381 agttccggct gacattagat tgacatccat caagtctaca actctgagag ttgaccagtc
     1441 gattcttaca ggtgaatctg tctcggtcat caagcatact gaccctgtcc ctgacccacg
     1501 agctgttaat caagacaaaa agaacatgct cttttctggc acaaacatcg ctgctggcaa
     1561 agctatggga gtggtggtgg cgactggagt caatactgag atcggcaaga tccgggatga
     1621 aatggttgca acagaacagg agagaacacc cctacagcag aagctggacg agtttgggga
     1681 acagctttcc aaagttatct ccctcatttg cattgcagtc tggatcatca acatcgggca
     1741 tttcaatgac ccagttcatg gtggctcttg gatcagaggt gccatctact actttaagat
     1801 tgcagtggcc ctggctgttg ctgccatccc tgagggtctg cctgctgtca tcaccacctg
     1861 cttggctctt ggaactcgaa ggatggcaaa gaaaaatgct attgttcgaa gtctgccttc
     1921 tgtggaaacc cttggttgta cttctgttat ctgctcagac aagaccggca cacttaccac
     1981 aaaccagatg tccgtctgca ggatgttcat tctggacaaa gtagaaggtg atacttgttc
     2041 ccttaatgag tttaccataa ctggatcaac ctatgcaccc attggagaag tgcaaaaaga
     2101 tgacaagcca gtgaaatgcc atcagtatga cgggcttgta gagttagcaa cgatctgtgc
     2161 tctgtgtaat gactctgctt tggattacaa tgaggcgaag ggtgtgtatg aaaaagttgg
     2221 agaagccaca gagactgctc tcacgtgcct ggtagagaag atgaatgtat tcgacacgga
     2281 gctgaaggga ctttctaaaa tagaacgcgc caacgcctgc aactcggtca taaaacagct
     2341 catgaagaag gaattcacgc tagagttttc acgtgataga aaatcaatgt ccgtctactg
     2401 tacaccaaac aaaccgagcc ggacgtccat gagcaagatg tttgtgaagg gtgctccaga
     2461 aggtgtcatc gacaggtgca cccacatccg agttggaagt accaaggtcc ccatgacgcc
     2521 tggtgttaaa cagaagatta tgtctgtcat tcgggagtgg ggcagtggca gcgacacact
     2581 gcggtgcctg gctctggcca ctcatgacaa cccgctgagg agagaggaga tgcacctgga
     2641 agattctgcg aacttcatca aatatgagac caatctgact ttcgttggct gtgtgggcat
     2701 gctggaccct cccaggattg aagtggcctc ttctgtgaag ctgtgccggc aagcgggcat
     2761 ccgagtcatc atgatcactg gggataacaa aggcactgct gtggccatct gtcgccgcat
     2821 tggcatcttt gggcaggatg aggatgtgac atcaaaggct tttacagggc gagaatttga
     2881 tgaattaagc ccctcagccc agagagacgc ctgcttaaat gcccgttgtt ttgctcgagt
     2941 tgaaccttcc cacaagtcta agatcgttga gttcctgcag tcctttgatg agatcacagc
     3001 tatgactggt gatggtgtga acgacgcgcc cgctctgaag aagtcggaaa tcgggattgc
     3061 catgggctca gggactgcag tggctaagac ggcctctgag atggtgctgg ccgacgacaa
     3121 cttctccacc atcgtggccg ctgttgagga ggggcgcgcc atctacaaca acatgaagca
     3181 gttcatccgc tacctcatct cctccaacgt gggggaggtg gtctgtatct tcctgacggc
     3241 agcccttggg tttcctgaag ctttgattcc tgtccagtta ctctgggtca atctggtgac
     3301 ggatggtctg cctgccactg cgctggggtt caatcctcca gatctggaca tcatgaacaa
     3361 acccccacgg aacccaaaag aaccgctgat cagcgggtgg ctctttttcc gttacctggc
     3421 tattggctgt tatgttggcg ctgccaccgt gggtgctgct gcgtggtggt tcatcgctgc
     3481 tgacggtggt ccgagagtct ccttctacca gctgagtcat ttcctgcagt gtaaggagga
     3541 caacccagac ttcgaaggag tggattgtgc aatctttgag tccccgtatc cgatgacaat
     3601 ggcactttct gttctagtca ccatagagat gtgcaatgcc ctcaacagct tgtctgaaaa
     3661 ccagtccctg ctgaggatgc ccccctggga gaacatctgg ctcgtgggct ccatctgctt
     3721 gtccatgtcc cttcacttct tgatcctcta cgtggaacct ttgccactca ttttccagat
     3781 cacaccgctg aatctgaccc agtggctgat ggtgctgaaa atctccctgc ctgtgatcct
     3841 catggacgag acgctcaagt ttgtggcccg aaactacctg gagcctgcaa tactggaggc
     3901 gaattcgcta gcgcttcgca ccggtgatat cgaacaaaaa ctaatatcag aggaagattt
     3961 ggaacagaaa ctaatcagtg aagaagattt agagcaaaag ttaatttccg aggaggactt
     4021 atagctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     4081 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     4141 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     4201 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     4261 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     4321 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4381 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4441 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4501 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4561 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4621 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4681 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4741 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4801 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4861 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4921 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4981 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5041 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5101 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     5161 tgaggatcgt ttcgcatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     5221 gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     5281 accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     5341 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     5401 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     5461 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     5521 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     5581 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     5641 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     5701 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     5761 aagcccggtg cctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5821 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5881 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5941 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6001 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6061 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     6121 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6181 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6241 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6301 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6361 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6421 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6481 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6541 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6601 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6661 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6721 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6781 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6841 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6901 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6961 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7021 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     7081 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7141 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7201 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7261 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7321 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7381 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7441 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7501 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7561 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7621 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7681 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7741 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7801 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7861 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7921 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7981 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8041 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8101 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8161 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8221 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     8281 cgggagatct cccgatcccc tatggt
//
